- Email: [email protected]
Novel reconstruction of the extrahepatic bile duct with a tissue-engineered bioabsorbable polymer graft
Vol. 221, No. 4S2, October 2015
Novel reconstruction of the extrahepatic bile duct with a tissue-engineered bioabsorbable polymer graft Xulong Zhu, MD, Qingdong Gao, MD, Weijie Yao, MD, Xiaopeng Yan, PhD, Junxi Xiang, MD, De Liu, MM, Zhen Qi Liu, MD, Yulong Song, MD, Yi Lv, Jianhui Li, MD Shaanxi Provincial People’s Hospital, Xi’an Jiaotong University, Xi’an, China INTRODUCTION: The purpose of this study was to evaluate the feasibility of using allogeneic porcine bone marrow mesenchymal stem cells (BMSCs) cultured onto a bioabsorbable polymer tube for the complete reconstruction of an extrahepatic bile duct in pigs. We also explored the potential benefit of local delivery of cytokines with biodegradable microspheres. METHODS: Porcine BMSCs were obtained by crista iliaca puncture from adult male pigs and cultured in vitro for 3 weeks. Cells at passage 2 were suspended in mixture of cytokines and Matrigel at a density of 2000000 cells/ml. The Matrigel solution was stratified on the bioabsorbable polymer tube to form a gel and cultured dynamically with differentiation medium for 1 week. Experimental models with common bile duct (CBD) defect were established and then CBD was reconstructed by duct to duct anastomosis with the polymer tube. Animals were relaparotomized at 1, 3 and 6 months after the grafting; subsequently, cholangiograms and monthly liver function tests were performed. We also conducted real-time qPCR analysis for the Y chromosome-specific gene sry in sorted recipient and donor derived cells from the same pig. RESULTS: All 18 recipient pigs survived until killing. Histological examination revealed incomplete epithelialization of the neo-bile duct at 1 month and 3 months after transplantation. At 6 months, the graft site was indistinguishable from the native duct. Real-time qPCR analysis for a donor specific gene (sry) in sorted CK19recipient cells suggested that fusion was a rare event in this experimental model. CONCLUSIONS: This newly designed substitute has potential for application as a novel treatment for hepatobiliary diseases. Osteopontin is a novel surrogate marker of SOX9-positive cancer stem cells in human hepatocellular carcinoma Takayuki Kawai, MD, Kentaro Yasuchika, Takamichi Ishii, Hokahiro Katayama, Satoshi Ogiso, MD, Sadahiko Kita, MD, Ken Fukumitsu, Etsuro Hatano, MD, PhD, Shinji Uemoto, MD, PhD Department of Surgery, Kyoto University, Kyoto, Japan INTRODUCTION: Although several markers of cancer stem cells (CSCs) have been reported in hepatocellular carcinoma (HCC), the HCC-CSC marker with useful surrogate marker is not identified. We aimed to determine if sex determining region Y-box 9 (SOX9), an important marker in normal liver development, can use as a new HCC-CSC marker, and if osteopontin can be a surrogate marker of SOX9. METHODS: We transfected the SOX9 promoter-driven enhanced green fluorescence protein gene into HCC cell lines, and investigated
Scientific Poster Presentations: 2015 Clinical Congress
e101
fluorescence-activated cell sorting (FACS)-isolated SOX9+/SOX9 cells. We examined SOX9 expression of consecutive 166 human surgical HCC specimens using immunohistochemistry, and analyzed serum osteopontin level of SOX9+/SOX9 patients. RESULTS: FACS-isolated single SOX9+ cells showed the ability to self-renew and differentiate into SOX9 cells, while single SOX9 cells never produced SOX9+ cells. SOX9+ cells displayed significantly greater proliferation capacity, higher sphere forming ability in vitro. Xenotransplantation into immunodeficiency mice revealed that SOX9+ cells could reproduce themselves, differentiate into SOX9 cells, and generate larger tumors in vivo. SOX9+ cells were found to be involved in EMT and the activation of TGFb/ Smad signaling. SOX9 expression was surrogated by osteopontin expression, which was suppressed by SOX9 knockdown and rescued by SOX9 overexpression. Immunohistochemistry of HCC specimens and measurement of serum osteopontin showed that compared to SOX9 patients, SOX9+ patients had significantly poorer recurrence-free survival, stronger venous invasion, and higher tumor/serum osteopontin expression. CONCLUSIONS: SOX9 is a new HCC-CSC marker correlated with EMT and TGFb/Smad signaling. Osteopontin should be expected the clinical application as a useful surrogate marker of SOX9. Pancreatoduodenectomy: improved access and quality with dedicated hepatopancreatobiliary surgical care in a community healthcare system Kerri A Simo, MD, Roberta Redfern, PhD, Chelsey Holbert, Joseph J Sferra, MD, FACS ProMedica Heath System, Toledo, OH INTRODUCTION: Recent studies note the majority of the United States remains underserved in hepatopancreatobiliary (HPB) surgical care. It has also been reported that pancreatoduodenectomy (PD) should be performed at high-volume centers by high-volume surgeons. METHODS: Analysis was performed to determine the effect of dedicated HPB surgical care in a tertiary hospital (TH) on access and surgical outcomes for PD in a community healthcare system (CHS). Hospital type (tertiary vs non-tertiary) and the effect of the addition of a fellowship trained HPB surgeon on PD access and quality were examined over a 5 year time period (9/1/09-8/ 31/14). RESULTS: 64 PDs performed at two hospitals by four surgeons during this time. Majority of PDs were performed at TH (n¼48; 75%). Percentage of patients with complications (44 vs 94%) and severity of complications were less at the TH (Clavien-Dindo Grade IV; p<0.0001). Lower 30- and 90-day mortality (both 4.1 vs 18.75%, p¼0.08) and decreased LOS (12.256 vs 20.77, p¼0.005) were also seen at the TH. Number of PDs/year increased 445% (from mean¼8.75/year to 36/year) following the addition of dedicated HPB surgical care. Decrease in patient migration to centers outside of the CHS was seen
Novel reconstruction of the extrahepatic bile duct with a tissue-engineered bioabsorbable polymer graft Xulong Zhu, MD, Qingdong Gao, MD, Weijie Yao, MD, Xiaopeng Yan, PhD, Junxi Xiang, MD, De Liu, MM, Zhen Qi Liu, MD, Yulong Song, MD, Yi Lv, Jianhui Li, MD Shaanxi Provincial People’s Hospital, Xi’an Jiaotong University, Xi’an, China INTRODUCTION: The purpose of this study was to evaluate the feasibility of using allogeneic porcine bone marrow mesenchymal stem cells (BMSCs) cultured onto a bioabsorbable polymer tube for the complete reconstruction of an extrahepatic bile duct in pigs. We also explored the potential benefit of local delivery of cytokines with biodegradable microspheres. METHODS: Porcine BMSCs were obtained by crista iliaca puncture from adult male pigs and cultured in vitro for 3 weeks. Cells at passage 2 were suspended in mixture of cytokines and Matrigel at a density of 2000000 cells/ml. The Matrigel solution was stratified on the bioabsorbable polymer tube to form a gel and cultured dynamically with differentiation medium for 1 week. Experimental models with common bile duct (CBD) defect were established and then CBD was reconstructed by duct to duct anastomosis with the polymer tube. Animals were relaparotomized at 1, 3 and 6 months after the grafting; subsequently, cholangiograms and monthly liver function tests were performed. We also conducted real-time qPCR analysis for the Y chromosome-specific gene sry in sorted recipient and donor derived cells from the same pig. RESULTS: All 18 recipient pigs survived until killing. Histological examination revealed incomplete epithelialization of the neo-bile duct at 1 month and 3 months after transplantation. At 6 months, the graft site was indistinguishable from the native duct. Real-time qPCR analysis for a donor specific gene (sry) in sorted CK19recipient cells suggested that fusion was a rare event in this experimental model. CONCLUSIONS: This newly designed substitute has potential for application as a novel treatment for hepatobiliary diseases. Osteopontin is a novel surrogate marker of SOX9-positive cancer stem cells in human hepatocellular carcinoma Takayuki Kawai, MD, Kentaro Yasuchika, Takamichi Ishii, Hokahiro Katayama, Satoshi Ogiso, MD, Sadahiko Kita, MD, Ken Fukumitsu, Etsuro Hatano, MD, PhD, Shinji Uemoto, MD, PhD Department of Surgery, Kyoto University, Kyoto, Japan INTRODUCTION: Although several markers of cancer stem cells (CSCs) have been reported in hepatocellular carcinoma (HCC), the HCC-CSC marker with useful surrogate marker is not identified. We aimed to determine if sex determining region Y-box 9 (SOX9), an important marker in normal liver development, can use as a new HCC-CSC marker, and if osteopontin can be a surrogate marker of SOX9. METHODS: We transfected the SOX9 promoter-driven enhanced green fluorescence protein gene into HCC cell lines, and investigated
Scientific Poster Presentations: 2015 Clinical Congress
e101
fluorescence-activated cell sorting (FACS)-isolated SOX9+/SOX9 cells. We examined SOX9 expression of consecutive 166 human surgical HCC specimens using immunohistochemistry, and analyzed serum osteopontin level of SOX9+/SOX9 patients. RESULTS: FACS-isolated single SOX9+ cells showed the ability to self-renew and differentiate into SOX9 cells, while single SOX9 cells never produced SOX9+ cells. SOX9+ cells displayed significantly greater proliferation capacity, higher sphere forming ability in vitro. Xenotransplantation into immunodeficiency mice revealed that SOX9+ cells could reproduce themselves, differentiate into SOX9 cells, and generate larger tumors in vivo. SOX9+ cells were found to be involved in EMT and the activation of TGFb/ Smad signaling. SOX9 expression was surrogated by osteopontin expression, which was suppressed by SOX9 knockdown and rescued by SOX9 overexpression. Immunohistochemistry of HCC specimens and measurement of serum osteopontin showed that compared to SOX9 patients, SOX9+ patients had significantly poorer recurrence-free survival, stronger venous invasion, and higher tumor/serum osteopontin expression. CONCLUSIONS: SOX9 is a new HCC-CSC marker correlated with EMT and TGFb/Smad signaling. Osteopontin should be expected the clinical application as a useful surrogate marker of SOX9. Pancreatoduodenectomy: improved access and quality with dedicated hepatopancreatobiliary surgical care in a community healthcare system Kerri A Simo, MD, Roberta Redfern, PhD, Chelsey Holbert, Joseph J Sferra, MD, FACS ProMedica Heath System, Toledo, OH INTRODUCTION: Recent studies note the majority of the United States remains underserved in hepatopancreatobiliary (HPB) surgical care. It has also been reported that pancreatoduodenectomy (PD) should be performed at high-volume centers by high-volume surgeons. METHODS: Analysis was performed to determine the effect of dedicated HPB surgical care in a tertiary hospital (TH) on access and surgical outcomes for PD in a community healthcare system (CHS). Hospital type (tertiary vs non-tertiary) and the effect of the addition of a fellowship trained HPB surgeon on PD access and quality were examined over a 5 year time period (9/1/09-8/ 31/14). RESULTS: 64 PDs performed at two hospitals by four surgeons during this time. Majority of PDs were performed at TH (n¼48; 75%). Percentage of patients with complications (44 vs 94%) and severity of complications were less at the TH (Clavien-Dindo Grade IV; p<0.0001). Lower 30- and 90-day mortality (both 4.1 vs 18.75%, p¼0.08) and decreased LOS (12.256 vs 20.77, p¼0.005) were also seen at the TH. Number of PDs/year increased 445% (from mean¼8.75/year to 36/year) following the addition of dedicated HPB surgical care. Decrease in patient migration to centers outside of the CHS was seen